Extraction and Isolation of Tyrosine Ammonia Lyase (TAL) and Diosgenin from <i>Persea americana</i> as Potent Inhibitors of Melanin Synthesis Involving <i>In Vivo</i> Study on Zebrafish Embryos

Tyrosine ammonia lyase is an enzyme in the natural phenol biosynthesis pathway. L-tyrosine is deaminated to p-Coumaric acid with the help of the enzyme tyrosine ammonia lyase. Tyrosine Ammonia Lyase (TAL) is extracted from the leaves of Persea americana. Diosgenin involved in the inhibition of intracellular tyrosinase is extracted along with the enzyme Tyrosine Ammonia Lyase from leaves of Persea americana, significantly increasing the activity of Tyrosine Ammonia Lyase. Diosgenin in the current study on analysis is highly increasing the activity of enzymes. The ability of the enzyme along with diosgenin to increase the activity of the enzyme is analysed in the current study. The enzyme along with diosgenin is used for depigmentation of skin. The methodology of protein separation on treatment with acetone was utilized for the extraction of the enzyme. p-Coumaric acid formation detected using a UV-Visible spectrophotometer at 380nm confirmed the presence of the enzyme. Defatting followed by microwave-assisted extraction and ultrasound-assisted extraction methodology were used for the extraction of diosgenin from leaves of Persea americana. The role of diosgenin in increasing the activity of the enzyme was carried out by analysing the enzyme activity at 410nm. The enzyme-activating ability of diosgenin was compared with that of magnesium sulphate-purified Tyrosine Ammonia Lyase enzyme and sodium sulphate-purified Tyrosine Ammonia Lyase enzyme. Zebrafish embryos were used as model organisms for the conduction of in vivo studies. kojic acid was used as a control in the in vivo studies to compare the effect of the enzyme extracted along with the diosgenin with that of kojic acid.